MTT法和CCK-8法检测中药抗病毒活性成分细胞毒性的比较

The comparision of MTT and CCK-8 by detecting cytotoxicity of antiviral active components in traditional Chinese medicine

中药抗病毒活性成分的筛选首先需要考察化合物在体外的细胞毒性.分别用MTT法和CCK-8法考察穿心莲内酯和脱水穿心莲内酯这两种存在于中药穿心莲中具有抗病毒潜力的成分在人肺腺癌A549细胞株的毒性情况.结果表明,CCK-8和MTT两种方法检测得到的细胞存活率的数据具有一致性,均提示在A549细胞株中,穿心莲内酯和脱水穿心内酯的无毒浓度分别在0 5μg/mL及0 10μg/mL之间.用MTT法测定穿心莲内酯和脱水穿心莲内酯的CC50值的变异系数(CV)均大于CCK-8法得到CV,表明CCK-8法检测的精密度较MTT法更高.回归方程的拟合优度的判定系数和细胞存活率的数据标准偏差数值表明CCK-8法测定的准确度相对MTT法更高.另外,CCK-8法操作更为简便和快速,对实验者和环境危害低.因而,CCK-8法值得在细胞增殖和药物毒性检测等实验中推广应用.

To investigate the cytotoxicity in vitro is the first step of screening the antiviral active oftraditional Chinese medicine(TCM).In our study,MTT and CCK-8 were used to test the cytotoxicity of twopromising antiviral components in a TCM namedAndrographis paniculata,andrographolide and 14-deoxy-11,12-didehydroandrographolide in human lung epithelial A549 cell line.Cell viabilities from the two methodswere identical that non-toxic concentrations of andrographolide and 14-deoxy-11,12-didehydroandrographolidewere 0—5 μg/mL and 0—10 μg/mL,respectively.Coefficient of variation(CV) of CC50 of andrographolideand 14-deoxy-11,12-didehydroandrographolide by MTT were both higher than that by CCK-8,which suggestedthat precision of CCK-8 was higher than MTT.Determination coefficients of goodness of fit in regressionequation and standard deviation of cell viability indicated that accuracy of CCK-8 was also higher than MTT.Furthermore,CCK-8 was easier and faster in manipulation and less dangerous to experimenter andenvironment,which was worthy applying in the test of cellular proliferation and toxicity of medicine.